Metabolite profiling reveals YihU as a novel hydroxybutyrate dehydrogenase for alternative succinic semialdehyde metabolism in Escherichia coli

Natsumi Saito, Martin Robert, Hayataro Kochi, Goh Matsuo, Yuji Kakazu, Tomoyoshi Soga, Masaru Tomita

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

The search for novel enzymes and enzymatic activities is important to map out all metabolic activities and reveal cellular metabolic processes in a more exhaustive manner. Here we present biochemical and physiological evidence for the function of the uncharacterized protein YihU in Escherichia coli using metabolite profiling by capillary electrophoresis time-of-flight mass spectrometry. To detect enzymatic activity and simultaneously identify possible substrates and products of the putative enzyme, we profiled a complex mixture of metabolites in the presence or absence of YihU. In this manner, succinic semialdehyde was identified as a substrate for YihU. The purified YihU Protein catalyzed in vitro the NADH-dependent reduction of succinic semialdehyde to γ-hydroxybutyrate. Moreover, a yihU deletion mutant displayed reduced tolerance to the cytotoxic effects of exogenous addition of succinic semialdehyde. Profiling of intracellular metabolites following treatment of E. coli with succinic semialdehyde supports the existence of a YihU-catalyzed reduction of succinic semialdehyde to γ-hydroxybutyrate in addition to its known oxidation to succinate and through the tricarboxylic acid cycle. These findings suggest that YihU is a novel γ-hydroxybutyrate dehydrogenase involved in the metabolism of succinic semialdehyde, and other potentially toxic intermediates that may accumulate under stress conditions in E. coli.

Original languageEnglish
Pages (from-to)16442-16451
Number of pages10
JournalJournal of Biological Chemistry
Volume284
Issue number24
DOIs
Publication statusPublished - 2009 Jun 12

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Hydroxybutyrate Dehydrogenase
Metabolites
Metabolism
Escherichia coli
Hydroxybutyrates
Capillary electrophoresis
Citric Acid Cycle
Poisons
Capillary Electrophoresis
Succinic Acid
Substrates
Enzymes
Complex Mixtures
NAD
Mass spectrometry
succinic semialdehyde
Mass Spectrometry
Proteins
Oxidation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Metabolite profiling reveals YihU as a novel hydroxybutyrate dehydrogenase for alternative succinic semialdehyde metabolism in Escherichia coli. / Saito, Natsumi; Robert, Martin; Kochi, Hayataro; Matsuo, Goh; Kakazu, Yuji; Soga, Tomoyoshi; Tomita, Masaru.

In: Journal of Biological Chemistry, Vol. 284, No. 24, 12.06.2009, p. 16442-16451.

Research output: Contribution to journalArticle

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